Monoacylglycerols for use in conjunction with a lipase inhibitor and/or diets low in fat and/or calories

ABSTRACT

In an embodiment, compositions comprising monoacylglycerols (MAG), such as sn-1(3) MAG, are administered with a lipase inhibitor, such as tetrahydrolipstatin, and/or with a diet low in fat and/or calories. In another embodiment, compositions comprising MAG, such as sn-1(3) MAG, are administered concurrently with a lipase inhibitor, such as tetrahydrolipstatin, and/or with fat-soluble nutrients. The compositions and the methods of using the compositions enhance absorption of fatty acids and fat-soluble nutrients such as fat-soluble vitamins and carotenoids to address nutritional deficiencies due to a weight loss diet used in association with a lipase inhibitor.

CROSS REFERENCE TO RELATED APPLICATIONS

The present application is a National Stage of International ApplicationNo. PCT/EP2014/073024, filed on Oct. 27, 2014, which claims priority toU.S. Provisional Patent Application No. 61/896,530, filed Oct. 28, 2013,the entire contents of which are being incorporated herein by reference.

BACKGROUND

The present disclosure generally relates to health and nutrition. Morespecifically, the present disclosure relates to nutritional compositionsthat can promote absorption of fatty acids and fat-soluble nutrients.

Gastric and pancreatic lipase inhibitors such as tetrahydrolipstatin(commercially available as XENICAL® and ALLI®) can be used asanti-obesity drugs because these compounds prevent fats from beinghydrolyzed and absorbed, thus decreasing calorie intake. Specifically,tetrahydrolipstatin binds covalently to the active site of such lipasesto block lipid hydrolysis.

In conjunction with changes in diet and lifestyle, a lipase inhibitorhas a higher efficacy than changes in diet and lifestyle alone. Finer etal., Int. J. Obes. Relat. Metab. Disord., 24(3):306-13 (March 2000).However, common side effects can include gastrointestinal problems, suchas oily stools; deficiencies in fat-soluble nutrients, includingvitamins A, D, E, K and beta-carotene; and liver or kidney damage.

Decreased lipid absorption can result in reduced absorption oflong-chain polyunsaturated fatty acids (LC-PUFAs). LC-PUFAs areimportant fatty acids obtained from the diet and needed for normal bodyfunction. Two critical LC-PUFAs, eicosapentaenoic acid (EPA) anddocosahexaenoic acid (DHA), are obtained from conversion ofalpha-linolenic acid (ALA) in the body, but this conversion occurs at avery low rate; thus supplementation of EPA and DHA is highlyrecommended. Therefore, the low LC-PUFA levels resulting from decreasedlipid absorption associated with lipase inhibitors or low fat diets canincrease the risk of cardiovascular and metabolic diseases, especiallyin cases where n-3 LC-PUFAs such as EPA and DHA are minimally absorbed.

Reduced absorption of fat-soluble nutrients, such as vitamins A, D, Eand K and carotenoids such as beta-carotene and lutein, can also resultfrom decreased lipid absorption. The resultant low levels of thesevitamins and carotenoids can elevate the risks of a variety ofconditions, such as cardiovascular diseases, coagulation disorders,impaired wound healing and immune function, and bone and muscle functiondisorders.

SUMMARY

The present disclosure provides compositions comprisingmonoacylglycerols (MAG), such as sn-1(3) MAG, for administration with alipase inhibitor, such as tetrahydrolipstatin, and/or with a diet low infat and/or calories. In sn-1(3) MAG, the sn-1 or sn-3 position isoccupied by an acyl group, such as a fatty acid, and the sn-2 positionis not occupied by a fatty acid.

Accordingly, in a general embodiment, the present disclosure provides amethod that includes administering a therapeutically effective amount ofmonoacylglycerols to an individual that has a characteristic selectedfrom the group consisting of periodic consumption of a lipase inhibitor,a diet low in fat, a diet low in calories, and combinations thereof.

In an embodiment, the monoacylglycerols comprise a therapeuticallyeffective amount of sn-1(3) monoacylglycerols.

In an embodiment, the amount of monoacylglycerols is therapeuticallyeffective to promote absorption of fatty acids and enhance delivery offat-soluble nutrients in the individual.

In an embodiment, the lipase inhibitor is tetrahydrolipstatin.

In an embodiment, the monoacylglycerols comprise a functional fattyacid.

In an embodiment, the method includes administering a fat-solublenutrient to the individual. The fat-soluble nutrient can be selectedfrom the group consisting of fat-soluble vitamins and carotenoids.

In an embodiment, the method includes administering themonoacylglycerols daily for at least three weeks.

In an embodiment, the monoacylglycerols include at least one of MAG-DHA,MAG-EPA or MAG-ARA.

In an embodiment a method is provided for treating a nutrient deficiencyin an individual that has a characteristic selected from the groupconsisting of periodic consumption of a lipase inhibitor, a diet low infat, a diet low in calories, and combinations thereof, comprisingadministering to an individual in need thereof a therapeuticallyeffective amount of a composition comprising monoacylglycerols, and afat-soluble nutrient, wherein the acyl group of the monoacylglycerols isselected from the group consisting of polyunsaturated fatty acids.

In another embodiment, a kit is provided. The kit includes: a firstcomposition comprising monoacylglycerols; a second compositioncomprising a lipase inhibitor; and a third composition that is anutritional composition that provides complete nutrition.

In an embodiment, the nutritional composition is designed such that apredetermined number of servings of the nutritional composition per dayprovide a diet low in at least one of fat or calories.

In another embodiment, a kit is provided. The kit includes: a firstcomposition comprising monoacylglycerols; a second compositioncomprising a lipase inhibitor; and a third composition that comprisesfat-soluble nutrients.

In an embodiment, the fat-soluble nutrients are selected from the groupconsisting of vitamin A, isoforms of vitamin A, vitamin D, isoforms ofvitamin D, vitamin E, isoforms of vitamin E, vitamin K, isoforms ofvitamin K, beta-carotene, lutein, and combinations thereof.

In another embodiment, a composition is provided. The compositionincludes a lipase inhibitor and sn-1(3) monoacylglycerols.

In an embodiment, the lipase inhibitor is tetrahydrolipstatin.

In an embodiment, the composition further includes fat-solublenutrients.

In an embodiment, the sn-1(3) monoacylglycerols include at least one ofMAG-DHA, MAG-EPA or MAG-ARA.

In an embodiment, no more than 25 weight %, preferably no more than 15weight %, of the total monoacylglycerols are Sn-2 monoacylglycerols.

An advantage of the present disclosure is to address side effects oflipase inhibitors, such as tetrahydrolipstatin, and/or diets low in fatand/or calories.

A further advantage of the present disclosure is to provide a foodcomposition that enables the efficient uptake of fatty acids despiteconditions of lipid malabsorption, such as lipase inhibitoradministration and diets low in calories and/or fat.

Another advantage of the present disclosure is to provide an optimalglyceride structure for substantial uptake of fatty acids despiteconditions of lipid malabsorption, such as lipase inhibitoradministration and diets low in calories and/or fat.

Still another advantage of the present disclosure is to enhanceabsorption of anti-inflammatory fatty acids despite conditions of lipidmalabsorption, such as lipase inhibitor administration and diets low incalories and/or fat.

An additional advantage of the present disclosure is to provide fattyacids in a form that do not need to be hydrolyzed prior to absorption.

Another advantage of the present disclosure is to enhance absorption offat-soluble nutrients despite conditions of lipid malabsorption, such aslipase inhibitor administration and diets low in calories and/or fat.

Still another advantage of the present disclosure is to provide fattyacids and enhance absorption of fat-soluble nutrients with compoundsthat have intrinsic emulsifying properties.

Yet another advantage of the present disclosure is to provide fattyacids and fat-soluble nutrients in a way that is compatible with a dietlow in calories and/or fat.

An additional advantage of the present disclosure is to provide fattyacids and fat-soluble nutrients in way that is well tolerated, withoutaggravating any steatorrhea.

Another advantage of the present disclosure is to correct nutritionaldeficiencies in LC-PUFAs and fat-soluble nutrients due to a weight lossdiet used in association with a lipase inhibitor.

Still another advantage of the present disclosure is to additionallyprovide preventive benefits with respect to cardiovascular and metabolicdisease.

Yet another advantage of the present disclosure is to provide kits thatenable a consumer to easily consume each component of the diet plan.

Additional features and advantages are described herein, and will beapparent from, the following Detailed Description and the Figures.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows the chemical structure of a sn-1(3) MAG. R is a fatty acid.

FIG. 2 depicts a graph showing the incorporation of EPA in red bloodcells resulting from treatments of control rats fed fish oil with orwithout tetrahydrolipstatin and rats fed tetrahydrolipstatin andvanillin acetal of 2-EPA (Group A), 1,3 diacetyl-2 EPA (Group B), and1,3 MAG-EPA (values are means±SEM, n=6).

FIG. 3 shows the timeline of a clinical study supporting the concept ofadministering sn-1(3) MAG to promote absorption of fatty acids andfat-soluble nutrients in a diet low in fat and/or calories inassociation with a lipase inhibitor.

FIG. 4 shows acute effects in the clinical study, namely pharmacokineticresults as measured by EPA in chylomicrons, AUC over 10 hourspostprandial.

FIG. 5 shows chronic effects in the clinical study, namely accretion ofEPA in erythrocytes as percentage of total fatty acids after 21 days oftreatment.

FIG. 6 shows chronic effects in the clinical study, namely accretion ofplasma in erythrocytes as percentage of total fatty acids after 21 daysof treatment.

FIG. 7 shows the chemical structure of tetrahydrolipstatin.

FIG. 8 shows the amount of β-carotene measured in the digestion solutionand in the micellar fraction (on cells) in an in-vitro digestion model.

DETAILED DESCRIPTION

All percentages expressed herein are by weight of the total weight ofthe composition unless expressed otherwise. When reference is made tothe pH, values correspond to pH measured at 25° C. with standardequipment. As used in this disclosure and the appended claims, thesingular forms “a,” “an” and “the” include plural referents unless thecontext clearly dictates otherwise. As used herein, “about” isunderstood to refer to numbers in a range of numerals. Moreover, allnumerical ranges herein should be understood to include all integers,whole or fractions, within the range. The composition disclosed hereinmay lack any element that is not specifically disclosed herein. Thus,the disclosure of an embodiment using the term “comprising” includes adisclosure of an embodiment “consisting essentially of” and anembodiment “consisting of” the referenced components. Any embodimentdisclosed herein can be combined with any other embodiment disclosedherein.

“Overweight” is defined for a human as a body mass index (BMI) between25 and 30. “Obese” is defined for a human as a BMI greater than 30. BMIis defined as the value resulting from the division equation in whichthe numerator is the weight in kilograms and the denominator is theheight in meters, squared.

“Prevention” includes reduction of risk and/or severity of a conditionor disorder. The terms “treatment,” “treat” and “to alleviate” includeboth prophylactic or preventive treatment (that prevent and/or slow thedevelopment of a targeted pathologic condition or disorder) andcurative, therapeutic or disease-modifying treatment, includingtherapeutic measures that cure, slow down, lessen symptoms of, and/orhalt progression of a diagnosed pathologic condition or disorder; andtreatment of patients at risk of contracting a disease or suspected tohave contracted a disease, as well as patients who are ill or have beendiagnosed as suffering from a disease or medical condition. The termdoes not necessarily imply that a subject is treated until totalrecovery. The terms “treatment” and “treat” also refer to themaintenance and/or promotion of health in an individual not sufferingfrom a disease but who may be susceptible to the development of anunhealthy condition. The terms “treatment,” “treat” and “to alleviate”are also intended to include the potentiation or otherwise enhancementof one or more primary prophylactic or therapeutic measures. The terms“treatment,” “treat” and “alleviate” are further intended to include thedietary management of a disease or condition or the dietary managementfor prophylaxis or prevention a disease or condition. A treatment can bepatient- or doctor-related.

As used herein, a “therapeutically effective amount” is an amount thatprevents a deficiency, treats a disease or medical condition in anindividual or, more generally, reduces symptoms, manages progression ofthe diseases or provides a nutritional, physiological, or medicalbenefit to the individual. The therapeutically effective amount that isrequired to achieve a therapeutic effect will, of course, vary with theparticular composition, the route of administration, the age and thecondition of the recipient, and the particular disorder or disease beingtreated.

“Animal” includes, but is not limited to, mammals, which includes but isnot limited to, rodents, aquatic mammals, domestic animals such as dogsand cats, farm animals such as sheep, pigs, cows and horses, and humans.Where “animal,” “mammal” or a plural thereof is used, these terms alsoapply to any animal that is capable of the effect exhibited or intendedto be exhibited by the context of the passage. As used herein, the term“patient” is understood to include an animal, especially a mammal, andmore especially a human that is receiving or intended to receivetreatment, as treatment is herein defined. While the terms “individual”and “patient” are often used herein to refer to a human, the presentdisclosure is not so limited. Accordingly, the terms “individual” and“patient” refer to any animal, mammal or human, having or at risk for amedical condition that can benefit from the treatment.

“Food product” and “food composition,” as used herein, are understood toinclude any number of optional additional ingredients, includingconventional food additives, for example one or more proteins,carbohydrates, fats, acidulants, thickeners, buffers or agents for pHadjustment, chelating agents, colorants, emulsifiers, excipients, flavoragents, minerals, osmotic agents, a pharmaceutically acceptable carrier,preservatives, stabilizers, sugars, sweeteners, texturizers and/orvitamins. The optional ingredients can be added in any suitable amount.

As used herein, “complete nutrition” includes nutritional products andcompositions that contain sufficient types and levels of macronutrients(protein, fats and carbohydrates) and micronutrients to be sufficient tobe a sole source of nutrition for the animal to which the composition isadministered. Patients can receive 100% of their nutritionalrequirements from such complete nutritional compositions. As usedherein, “incomplete nutrition” includes nutritional products orcompositions that do not contain sufficient levels of macronutrients(protein, fats and carbohydrates) or micronutrients to be sufficient tobe a sole source of nutrition for the animal to which the composition isadministered. Partial or incomplete nutritional compositions can be usedas a nutritional supplement.

A “kit” means that the components of the kit are physically associatedin or with one or more containers and considered a unit for manufacture,distribution, sale, or use. Containers include, but are not limited to,bags, boxes, cartons, bottles, packages of any type or design ormaterial, over-wrap, shrink-wrap, affixed components (e.g., stapled,adhered, or the like), or combinations thereof.

“Concurrent” and “concurrently” in the context of the present disclosuremean in the same day, preferably in the same twelve hour period, morepreferably within the same hour, most preferably simultaneously.

A “diet low in fat and/or calories” is a diet in which 1,000calories/day or less and/or 150 g of calories from fat/day or less areconsumed. In such a diet, preferably 800 calories/day or less areconsumed and preferably 125 g of calories from fat/day or less areconsumed.

The present disclosure provides compositions comprisingmonoacylglycerols (MAG), such as sn-1(3) MAG. FIG. 1 depicts thechemical structure of a sn-1(3) MAG, and R is the fatty acid. Forexample, 1,3 MAG-EPA is the chemical structure shown in FIG. 1 in whichR is EPA. The sn-1(3) MAG may be chemically synthesized, for exampleusing glycerol and fish oil. The composition can comprise the sn-1(3)MAG in an amount corresponding to 1% to 40% of the energy of thecomposition, preferably from 5% to 40% of the energy of the composition.

Preferably the compositions comprise MAG in an amount that istherapeutically effective for providing fatty acids and/or enhancingabsorption of fat-soluble nutrients. More preferably the compositionscomprise sn-1(3) MAG in an amount that is therapeutically effective forproviding fatty acids and/or enhancing absorption of fat-solublenutrients.

The present disclosure also provides methods in which such compositionsare administered concurrently with a lipase inhibitor, such astetrahydrolipstatin (FIG. 7), and/or with a diet low in fat and/orcalories. Preferably, the individual is consuming the lipase inhibitorperiodically, such as weekly or daily. Preferably, compositionscomprising a therapeutically effective amount of monoacylglycerols(MAG), such as sn-1(3) MAG, are administered daily and concurrently witha lipase inhibitor and/or with a diet low in fat and/or calories for atleast three weeks, more preferably at least eight weeks, and mostpreferably at least twelve weeks. A non-limiting example of a diet lowin fat and calories is a diet that solely uses liquid products designedto restrict caloric intake, while providing complete nutrition, such asthe full meal replacement liquid products of Nestle S.A.'s OPTIFAST®.For example, a diet low in fat and calories may be a diet in which fiveservings of a composition are consumed daily, and each serving has about160 calories, about 25 calories from fat, about 20 g of carbohydrates,and about 14 g of protein.

The present disclosure also provides methods in which compositionscomprising MAG are administered concurrently with a lipase inhibitor,such as tetrahydrolipstatin (FIG. 2), and/or with fat-soluble nutrients.Non-limiting examples of fat-soluble nutrients include fat-solublevitamins, such as vitamins A, D, E and K and their isoforms, andcarotenoids, such as beta-carotene and lutein. Preferably, theindividual is consuming the lipase inhibitor periodically, such asweekly or daily. Preferably, the compositions comprisingmonoacylglycerols (MAG), such as sn-1(3) MAG, are administered daily andconcurrently with the lipase inhibitor and/or with fat-soluble nutrientsfor at least three weeks, more preferably at least eight weeks, and mostpreferably at least twelve weeks.

Sn-1(3) MAG interacts with vitamins due to its emulsifying propertiesand thus facilitates absorption of vitamins A, D, E and K, potentiallysynergistically. Therefore the compositions according to the presentdisclosure can correct nutritional deficiencies in vitamins and fattyacids (PUFA), for example nutritional deficiencies in vitamins and fattyacids (PUFA) related to low fat diets. Preferably the lipase inhibitoris administered in an amount that is therapeutically effective todecrease the gastrointestinal absorption of fats.

According to the present disclosure, concurrent administration of thelipase inhibitor and the composition comprising MAG includesadministration of the lipase inhibitor separately from the compositioncomprising monoacylglycerols and also includes administration of thelipase inhibitor and the MAG in the same composition. Similarly,concurrent administration of the lipase inhibitor and the diet low infat and/or calories includes administration of the lipase inhibitorseparately from a serving of the diet low in fat and/or calories andalso includes administration of the lipase inhibitor and a serving ofthe diet low in fat and/or calories in the same composition. Alsosimilarly, concurrent administration of the lipase inhibitor and thefat-soluble nutrients includes administration of the lipase inhibitorseparately from a serving of the fat-soluble nutrients and also includesadministration of the lipase inhibitor and a serving of the fat-solublenutrients in the same composition.

The recipient of administration may be any individual but preferably isan overweight or obese individual or an individual at risk of overweightor obesity. The compositions comprising MAG can be implemented in amethod of treating or preventing overweight or obesity. In someembodiments, the compositions comprising MAG can provide n-3 LC-PUFAs inan amount that is therapeutically effective to treat or preventcardiovascular disease or rheumatoid arthritis; increase the level ofeicosanoids such as prostaglandin-3; enhance brain and retinadevelopment; treat or prevent vision decline; and/or enhance immunefunction.

The acyl group of the MAG may be a functional fatty acid. A functionalfatty acid is a fatty acid that provides a health benefit to anindividual administered the fatty acid. Non-limiting examples offunctional fatty acids include eicosapentaenoic acid (EPA),docosahexaenoic acid (DHA), α-linolenic acid (ALA), stearidonic acid(SA), γ-linolenic acid (GLA), dihomo-γ-linolenic acid (DGLA),docosapentanenoic acid (DPA), sciadonic acid, and juniperonic acid.Sciadonic acid is 5Z, 11Z, 14Z-eicosatrienoic acid. Juniperonic acid is5(Z), 11(Z), 14(Z), 17(Z)-eicosatetraenoic acid.

Non-limiting examples of MAG that may be used in the compositionsprovided by the present disclosure include:

sn-1(3)-monoeicosapentaenoylglycerol

sn-1(3)-monodocosahexaenoylglycerol

sn-1(3)-monooctadecatrienoylglycerol

sn-1(3)-monooctadecatetraenoylglycerol

sn-1(3)-monoeicosatrienoylglycerol

sn-1(3)-monodocosapentaenoylglycerol

sn-1(3)-monosciadonylglycerol

sn-1(3)-monojuniperonylglycerol

and combinations thereof.

Of course, the composition may comprise a mixture of different MAG withdifferent fatty acids in the sn-1(3) position. The fatty acids may bemixed to achieve a particular ratio between n-3 and n-6 fatty acids.Non-limiting examples of suitable n-3 fatty acids include α-linolenicacid, stearidonic acid, eicosatrienoic acid, n-3 eicosatetraenoic acid,eicosapentaenoic acid, clupanodonic acid, docosahexaenoic acid, n-3tetracosapentaenoic acid, and n-3 tetracosahexaenoic acid. Non-limitingexamples of suitable n-6 fatty acids include linoleic acid, γ-linolenicacid, n-6 eicosadienoic acid, dihomo-γ-linolenic acid, arachidonic acid,n-6 docosadienoic acid, adrenic acid, n-6 docosapentaenoic acid, andcalendic acid.

In an embodiment, the composition contains a combination of differentsn-1(3) MAG such that the ratio of n-3 to n-6 fatty acids is about 5:1to about 15:1, preferably about 8:1 to about 10:1.

Optionally, the composition contains sn-2 MAG in addition to the sn-1(3)MAG. Depending on the nature of the fatty acid used as acyl-group in thesn-1(3) position, such mixtures may form automatically throughisomerisation. Therefore, an embodiment of the composition comprises 25%or less by weight of the total MAG as sn-2 MAG, preferably 15% or lessby weight of the total MAG as sn-2 MAG. The sn-1 and sn-3 positions ofthe sn-2 MAG can be blocked by protective groups to limit isomerization.Non-limiting examples of suitable protective groups include acetylgroups, ethyl groups, propyl groups, vanillin, and other molecules ableto form acetals. In some embodiments, the protective group bridges thehydroxyl groups in sn-1 and sn-3 positions.

Non-limiting examples of suitable sn-2 MAG include:

1,3-diacetyl-2-eicosapentaenoylglycerol

1,3-diethyl-2-eicosapentaenoylglycerol

1,3-dipropyl-2-eicosapentaenoylglycerol

a vanillin derivative of sn-2 monoeicosapentaenoylglycerol

other acetal derivatives of monoeicosapentaenoylglycerol

and combinations thereof.

Unwanted isomerisation may be prevented or at least slowed significantlyby adjusting the pH to the neutral range and/or by keeping thetemperature of the composition low. Therefore, the composition may havea pH in the range of 5 to 8, preferably 5 to 7. The composition may bestored at 8° C. or below.

Isomerization of the MAG may further be prevented, even in the bodyafter consumption, by inhibiting the action of lipase B. Therefore, thecomposition may comprise a lipase B inhibitor. Lipase B inhibitors areknown to those of skill in the art. Edible lipase B inhibitors arepreferred. “Edible” means that a material is approved for human oranimal consumption.

The composition provided by the present disclosure may be any kind ofedible composition. Preferably, the composition is a composition to beadministered orally or enterally. For example, the composition may beselected from the group consisting of a food product, an animal foodproduct, a pharmaceutical composition, a nutritional composition, anutraceutical, a drink, a food additive, and a medicament. In anembodiment, the composition is a liquid nutritional formula to beadministered enterally, e.g., in hospitals.

In an embodiment, the composition is a powdered composition to bereconstituted in milk or water. If the composition has the form of apowder, the powder may be a shelf stable powder. Shelf stability can beobtained, for example, by providing the composition with a wateractivity less than 0.2, for example in the range of 0.05 to 0.19,preferably in the range of 0.05 to 0.15. Water activity (a_(w)) is ameasurement of the energy status of the water in a system and defined asthe vapor pressure of water divided by that of pure water at the sametemperature; therefore, pure distilled water has a water activity ofexactly one.

The composition comprising MAG may be a nutritional composition thatalso contains a protein source and/or a carbohydrate source. Easilydigestible carbohydrates and/or proteins are preferred. Proteins thatare hydrolyzed at least partially are easier to digest and absorb.Therefore, the protein may have a degree of hydrolysis between 2 and20%. If hydrolyzed proteins are required, the hydrolysis process may becarried out using any process known in the art. For example, a proteinhydrolysate may be prepared by enzymatically hydrolysing a proteinfraction in one or more steps. For an extensively hydrolysed protein,the proteins may be subjected to triple hydrolysis using Alcalase 2.4 L(EC 940459), then Neutrase 0.5 L (obtainable from Novo Nordisk FermentAG) and then pancreatin at 55° C.

The nutritional composition may be a source of complete nutrition or maybe a source of incomplete nutrition. As used herein, “completenutrition” includes nutritional products and compositions that containsufficient types and levels of macronutrients (protein, fats andcarbohydrates) and micronutrients to be sufficient to be a sole sourceof nutrition for the animal to which the composition is administered.Individuals can receive 100% of their nutritional requirements from suchcomplete nutritional compositions. As used herein, “incompletenutrition” includes nutritional products or compositions that do notcontain sufficient levels of macronutrients (protein, fats andcarbohydrates) or micronutrients to be sufficient to be a sole source ofnutrition for the animal to which the composition is administered.Partial or incomplete nutritional compositions can be used as anutritional supplement.

In an embodiment, the diet low in fat and/or calories is achieved byconsuming several compositions daily such that the compositions providedcomplete nutrition. The MAG may be provided by one or more of thesecompositions. As a non-limiting example, the diet low in fat and/orcalories may be achieved by consuming five compositions daily, and eachof the compositions can have 160 calories, 25 g of fat, sn-1(3) MAG, andoptionally sn-2 MAG. The lipase inhibitor, such as tetrahydrolipstatin,may also be consumed daily and thus concurrently to such compositions asdefined by the present disclosure.

To encourage concurrent administration, the composition comprising MAG,the servings of the diet low in fat and/or calories, and the lipaseinhibitor may be provided in a kit. For example, each of the compositioncomprising MAG, the servings of the diet low in fat and/or calories, andthe lipase inhibitor may be provided in separate containers in a singlepackage. Similarly, the composition comprising MAG, the fat-solublenutrients, and the lipase inhibitor may be provided in a kit. Forexample, each of the compositions comprising MAG, the fat-solublenutrients, and the lipase inhibitor may be provided in separatecontainers in a single package.

EXAMPLES

The following non-limiting examples present scientific data developingand supporting the concept of administering sn-1(3) MAG to promoteabsorption of fatty acids and fat-soluble nutrients in a diet low in fatand/or calories in association with a lipase inhibitor.

Example 1

The concept was tested in a lipid maldigestion or malabsorption ratmodel. The maldigestion or malabsorption condition was obtained usingXENICAL® (ORLISTAT), a pancreatic and gastric lipases inhibitor(tetrahydrolipstatin; see FIG. 7). Rats were fed during 21 days withlong-chain polyunsaturated fatty acid (LC-PUFA) supplements containingmainly eicosapentaenoic (EPA) acid. Fish oil was used as a source oftriacylglycerols, and different EPA glycerides were evaluated. XENICAL®(ORLISTAT) was given at a level sufficient to decrease lipid absorptionby 40%. A group receiving fish oil without XENICAL® (ORLISTAT) was usedas a positive control. At different time intervals (D3, D7, D14 andD21), the fatty acid profiles of red blood cell and plasma lipids weredetermined. At the end of the experiment, the fatty acid profiles ofdifferent tissues were determined.

The main objective was to follow the level of EPA in red blood cell andplasma lipids. The main comparison evaluated was the difference in EPAlevel between the group receiving EPA-containing sn-1(3) MAG (FIG. 1) incombination with XENICAL® (ORLISTAT) and the positive control group(fish oil+XENICAL® (ORLISTAT)).

As an example, data obtained for EPA levels in red blood cell lipids atday 7 are reported in FIG. 2. The statistical evaluation revealed thatthe use of XENICAL® (ORLISTAT) decreases EPA incorporation in red bloodcells (comparison between the group receiving fish oil in combinationwith XENICAL® (ORLISTAT) and the group receiving fish oil withoutXENICAL® (ORLISTAT)). This comparison corroborates the validity of themodel. The level of EPA incorporated in red blood cells in animalsreceiving the sn-1(3) MAG that contained EPA is statistically higherthat the fish oil+group receiving fish oil in combination with XENICAL®(ORLISTAT) (all P values lower than 0.05), and more surprisingly, evenhigher than the fish oil group.

This example clearly demonstrates that in conditions of lipidmaldigestion or malabsorption, the incorporation of LC-PUFAs provided astriacylglycerols is reduced. However, if LC-PUFAs are provided assn-1(3) MAG (Group C), the incorporation in tissue is improved, even inconditions of lipid maldigestion or malabsorption.

Example 2

This clinical study compared the efficacy of sn-1(3) MAG and fish oil(TAG) in delivering EPA in humans under lipid maldigestion conditionsinduced by XENICAL® (ORLISTAT). The comparison was tested in volunteershaving a BMI of 37-40 kg/m² and treated with XENICAL® (ORLISTAT) toinduce lipid maldigestion or not treated with XENICAL® (ORLISTAT). Theprimary objective was to assess accretion of EPA in erythrocytes over 21days when consumed as fish oil (TAG) or sn-1(3) MAG. The secondaryobjectives were to assess accretion of EPA in plasma over 21 days andalso to assess the pharmacokinetics of EPA after an acute dose either inthe form of sn-1(3) MAG or TAG (AUC in chylomicrons over 10 hourspostprandial). See FIG. 3.

TABLE 1 Experimental Groups Group Oil Type and Total EPA Orlistat ® (120mg) No. number per day (mg) and number per day 1 (n = 11) Fish oil 3 504No — 2 (n = 11) MAG 3 500 No — 3 (n = 11) Fish oil 3 504 Yes 3 4 (n =11) MAG 3 500 Yes 3

The pharmacokinetic results (FIG. 4) show that the acute effect fromtreatment with sn-1(3) MAG and XENICAL® (ORLISTAT) is statisticallysignificant relative to treatment with fish oil and XENICAL® (ORLISTAT)(p=0.0125). The accretion of EPA in erythrocytes after 21 days (FIG. 5)shows that the chronic effect of treatment with sn-1(3) MAG and XENICAL®(ORLISTAT) is statistically significant, especially in comparison totreatment with fish oil and XENICAL® (ORLISTAT) (p=0.0001). Theaccretion of EPA in plasma after 21 days (FIG. 6) shows that the chroniceffect of treatment with sn-1(3) MAG and XENICAL® (ORLISTAT) isstatistically significant relative to treatment with fish oil andXENICAL® (ORLISTAT) (p=0.0003).

This clinical trial confirmed that, in obese subjects treated withXENICAL® (ORLISTAT), sn-1(3) MAG is a better carrier for EPA than fishoil (TAG).

Example 3 In Vitro Digestion to Assess Lipidic ComponentsBioaccessibility

Simulated or in vitro digestion is a model to be used to assess thestability of lipidic components such as liposoluble vitamins andcarotenoids during the digestive phases (oral, gastric and smallintestinal) and the extent of partitioning of lipidic components intomixed bile salt micelle fraction (essential step for absorption oflipophiles). Partitioning of lipidic components into mixed bile saltmicelle is also referred as “bioaccessibility” and expressed asefficiency of micellarization. In each step type of enzymes are adaptedas needed (e.g malabsorption vs. control) as well enzymes that are fitto purpose (e.g., TAG, MAG, vitamins, carotenoids).

In the procedure fish oil was used as a source of triacylglycerols(TAG). Monoacylglycerols (MAG) were purchased from Cognis GmbH, Germanyand mixed with sunflower oil at the ratio of 1:0.8 (w/w).

Triacylglycerols and Monoacylglycerols mixed with 3-carotene wereassessed in vitro using a digestion model where digestive enzymes andbile salts were reduced to obtain mal-digestion and mal-absorptionconditions.

To simulate gastric digestion, MAG or TAG plus β-carotene were incubated10 min at 37 C, in order to have a homogeneous mixture. A solution ofdifferent salts (NaCl, CaCl2 and KCl) and 1% F-127 emulsifier wereadded. pH was adjusted to 2.5 before the addition of pepsin. Thedigestion solution was incubated 1 h at 37° C.

To mimic intestinal digestion, pH was adjusted to 6, bile extract,pancreatin and lipase solutions were added to the previous digestionsolutions. pH was adjusted to 6.5 and the simulated digestions incubatedfor 2 h at 37° C.

After a liquid/liquid extraction, levels of β-carotene were measured byHPLC under the following analytical conditions: mobile phase:acetonitrile/terahydrofurane/methanol/1% ammonium acetate, flow rate:1.5 mL/min. Detection: photodiode array and fluorometry.

The digestion of β-carotene was followed through the above mentioned invitro digestion system to evaluate whether β-carotene digestion was moreeffective with MAG than with TAG

In the above described in vitro model for mal-digestion andmal-absorption conditions, it was observed that MAG has the tendency toimprove digestion and micellization of β-carotene when compared to TAG(FIG. 8). From FIG. 8 is seen that the amount of β-carotene measured inthe digestion solution and in the micellar fraction (on cells) is higherwith MAG than TAG, indicating improved digestion and micellization ofβ-carotene.

It should be understood that various changes and modifications to thepresently preferred embodiments described herein will be apparent tothose skilled in the art. Such changes and modifications can be madewithout departing from the spirit and scope of the present subjectmatter and without diminishing its intended advantages. It is thereforeintended that such changes and modifications be covered by the appendedclaims.

The invention is claimed as follows:
 1. A method of improving absorptionof fat-soluble nutrients in an individual having a diet low in calories,the method comprising concurrently administering beta-carotene and atherapeutically effective amount of monoacylglycerols to the individual.2. The method of claim 1 wherein the monoacylglycerols comprise atherapeutically effective amount of sn-1(3) monoacylglycerols.
 3. Themethod of claim 1 wherein the amount of monoacylglycerols istherapeutically effective to promote absorption of fatty acids andenhance delivery of fat-soluble nutrients in the individual.
 4. Themethod of claim 1 wherein the monoacylglycerols comprise a functionalfatty acid.
 5. The method of claim 1 comprising administering anadditional fat-soluble nutrient to the individual.
 6. The method ofclaim 5 wherein the additional fat-soluble nutrient is selected from thegroup consisting of fat-soluble vitamins and carotenoids.
 7. The methodof claim 1 comprising administering the beta-carotene and themonoacylglycerols to the individual daily for at least three weeks. 8.The method of claim 1 wherein the monoacylglycerols (MAG) are selectedfrom the group consisting of MAG-DHA, MAG-EPA and MAG-ARA.
 9. The methodof claim 1 wherein the beta-carotene and the monoacylglycerols areprovided by a composition in which no more than 15 wt. % of the totalmonoacylglycerols are Sn-2 monoacylglycerols.
 10. The method of claim 9wherein the monoacylglycerols are 5% to 40% of the energy of thecomposition.
 11. The method of claim 1 wherein the diet low in caloriesis 800 calories/day or less consumed by the individual.
 12. The methodof claim 1 wherein the diet low in calories is a diet that solely usesliquid products restricting caloric intake while providing completenutrition.
 13. The method of claim 12 wherein the monoacylglycerols areprovided by one or more of the liquid products.
 14. The method of claim5 wherein the additional fat-soluble nutrient comprises at least onefat-soluble vitamin selected from the group consisting of vitamin A,vitamin D, vitamin E, vitamin K, and an isoform thereof.
 15. The methodof claim 5 wherein the additional fat-soluble nutrient comprises atleast one carotenoid selected from the group consisting of beta-caroteneand lutein.